Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Rapid production of new oligodendrocytes is required in the earliest stages of motor-skill learning


We identified mRNA encoding the ecto-enzyme Enpp6 as a marker of newly forming oligodendrocytes, and used Enpp6 in situ hybridization to track oligodendrocyte differentiation in adult mice as they learned a motor skill (running on a wheel with unevenly spaced rungs). Within just 2.5 h of exposure to the complex wheel, production of Enpp6-expressing immature oligodendrocytes was accelerated in subcortical white matter; within 4 h, it was accelerated in motor cortex. Conditional deletion of myelin regulatory factor (Myrf) in oligodendrocyte precursors blocked formation of new Enpp6+ oligodendrocytes and impaired learning within the same 2−3 h time frame. This very early requirement for oligodendrocytes suggests a direct and active role in learning, closely linked to synaptic strengthening. Running performance of normal mice continued to improve over the following week accompanied by secondary waves of oligodendrocyte precursor proliferation and differentiation. We concluded that new oligodendrocytes contribute to both early and late stages of motor skill learning.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type



Prices may be subject to local taxes which are calculated during checkout

Figure 1: Time course of motor skill learning in mice and the requirement for Myrf.
Figure 2: Oligodendrocyte dynamics during motor-skill learning.
Figure 3: Enpp6 marks newly forming oligodendrocytes.
Figure 4: Enpp6highMbp+ newly formed oligodendrocytes express myelin structural proteins and synthesize myelin.
Figure 5: Visualization of Enpp6high cells in the developing mouse forebrain by ISH.
Figure 6: Rapid increase in Enpp6high newly forming oligodendrocytes in response to motor-skill learning.
Figure 7: Increased production of Enpp6+ newly formed oligodendrocytes was a response to motor learning, not physical exercise.


  1. Sturrock, R.R. Myelination of the mouse corpus callosum. Neuropathol. Appl. Neurobiol. 6, 415–420 (1980).

    Article  CAS  Google Scholar 

  2. Yeung, M.S. et al. Dynamics of oligodendrocyte generation and myelination in the human brain. Cell 159, 766–774 (2014).

    Article  CAS  Google Scholar 

  3. Dimou, L., Simon, C., Kirchhoff, F., Takebayashi, H. & Götz, M. Progeny of Olig2-expressing progenitors in the gray and white matter of the adult mouse cerebral cortex. J. Neurosci. 28, 10434–10442 (2008).

    Article  CAS  Google Scholar 

  4. Rivers, L.E. et al. PDGFRA/NG2 glia generate myelinating oligodendrocytes and piriform projection neurons in adult mice. Nat. Neurosci. 11, 1392–1401 (2008).

    Article  CAS  Google Scholar 

  5. Lasiene, J., Matsui, A., Sawa, Y., Wong, F. & Horner, P.J. Age-related myelin dynamics revealed by increased oligodendrogenesis and short internodes. Aging Cell 8, 201–213 (2009).

    Article  CAS  Google Scholar 

  6. Kang, S.H., Fukaya, M., Yang, J.K., Rothstein, J.D. & Bergles, D.E. NG2+ CNS glial progenitors remain committed to the oligodendrocyte lineage in postnatal life and following neurodegeneration. Neuron 68, 668–681 (2010).

    Article  CAS  Google Scholar 

  7. Zhu, X. et al. Age-dependent fate and lineage restriction of single NG2 cells. Development 138, 745–753 (2011).

    Article  CAS  Google Scholar 

  8. Simon, C., Götz, M. & Dimou, L. Progenitors in the adult cerebral cortex: cell cycle properties and regulation by physiological stimuli and injury. Glia 59, 869–881 (2011).

    Article  Google Scholar 

  9. Young, K.M. et al. Oligodendrocyte dynamics in the healthy adult CNS: evidence for myelin remodeling. Neuron 77, 873–885 (2013).

    Article  CAS  Google Scholar 

  10. Emery, B. et al. Myelin gene regulatory factor is a critical transcriptional regulator required for CNS myelination. Cell 138, 172–185 (2009).

    Article  CAS  Google Scholar 

  11. Koenning, M. et al. Myelin gene regulatory factor is required for maintenance of myelin and mature oligodendrocyte identity in the adult CNS. J. Neurosci. 32, 12528–12542 (2012).

    Article  CAS  Google Scholar 

  12. Hornig, J. et al. The transcription factors Sox10 and Myrf define an essential regulatory network module in differentiating oligodendrocytes. PLoS Genet. 9, e1003907 (2013).

    Article  Google Scholar 

  13. McKenzie, I.A. et al. Motor skill learning requires active central myelination. Science 346, 318–322 (2014).

    Article  CAS  Google Scholar 

  14. Freeman, S.A. et al. Acceleration of conduction velocity linked to clustering of nodal components precedes myelination. Proc. Natl. Acad. Sci. USA 112, E321–E328 (2015).

    Article  CAS  Google Scholar 

  15. Fünfschilling, U. et al. Glycolytic oligodendrocytes maintain myelin and long-term axonal integrity. Nature 485, 517–521 (2012).

    Article  Google Scholar 

  16. Lee, Y. et al. Oligodendroglia metabolically support axons and contribute to neurodegeneration. Nature 487, 443–448 (2012).

    Article  CAS  Google Scholar 

  17. Volkmar, F.R. & Greenough, W.T. Rearing complexity affects branching of dendrites in the visual cortex of the rat. Science 176, 1445–1447 (1972).

    Article  CAS  Google Scholar 

  18. Bliss, T.V. & Collingridge, G.L. A synaptic model of memory: long-term potentiation in the hippocampus. Nature 361, 31–39 (1993).

    Article  CAS  Google Scholar 

  19. Milner, B., Squire, L.R. & Kandel, E.R. Cognitive neuroscience and the study of memory. Neuron 20, 445–468 (1998).

    Article  CAS  Google Scholar 

  20. Lee, K.J., Rhyu, I.J. & Pak, D.T. Synapses need coordination to learn motor skills. Rev. Neurosci. 25, 223–230 (2014).

    Article  Google Scholar 

  21. Xu, T. et al. Rapid formation and selective stabilization of synapses for enduring motor memories. Nature 462, 915–919 (2009).

    Article  CAS  Google Scholar 

  22. Li, Q., Brus-Ramer, M., Martin, J.H. & McDonald, J.W. Electrical stimulation of the medullary pyramid promotes proliferation and differentiation of oligodendrocyte progenitor cells in the corticospinal tract of the adult rat. Neurosci. Lett. 479, 128–133 (2010).

    Article  CAS  Google Scholar 

  23. Gibson, E.M. et al. Neuronal activity promotes oligodendrogenesis and adaptive myelination in the mammalian brain. Science 344, 1252304 (2014).

    Article  Google Scholar 

  24. Greiner-Tollersrud, L., Berg, T., Stensland, H.M., Evjen, G. & Greiner-Tollersrud, O.K. Bovine brain myelin glycerophosphocholine choline phosphodiesterase is an alkaline lysosphingomyelinase of the eNPP-family, regulated by lysosomal sorting. Neurochem. Res. 38, 300–310 (2013).

    Article  CAS  Google Scholar 

  25. Sakagami, H. et al. Biochemical and molecular characterization of a novel choline-specific glycerophosphodiester phosphodiesterase belonging to the nucleotide pyrophosphatase/phosphodiesterase family. J. Biol. Chem. 280, 23084–23093 (2005).

    Article  CAS  Google Scholar 

  26. Morita, J. et al. Structure and biological function of ENPP6, a choline-specific glycerophosphodiester-phosphodiesterase. Sci. Rep. 6, 20995 (2016).

    Article  CAS  Google Scholar 

  27. Zhang, Y. et al. An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex. J. Neurosci. 34, 11929–11947 (2014).

    Article  CAS  Google Scholar 

  28. Hughes, E.G., Kang, S.H., Fukaya, M. & Bergles, D.E. Oligodendrocyte progenitors balance growth with self-repulsion to achieve homeostasis in the adult brain. Nat. Neurosci. 16, 668–676 (2013).

    Article  CAS  Google Scholar 

  29. Walker, M.P., Brakefield, T., Morgan, A., Hobson, J.A. & Stickgold, R. Practice with sleep makes perfect: sleep-dependent motor skill learning. Neuron 35, 205–211 (2002).

    Article  CAS  Google Scholar 

  30. Trapp, B.D., Nishiyama, A., Cheng, D. & Macklin, W. Differentiation and death of premyelinating oligodendrocytes in developing rodent brain. J. Cell Biol. 137, 459–468 (1997).

    Article  CAS  Google Scholar 

  31. Chang, A., Tourtellotte, W.W., Rudick, R. & Trapp, B.D. Premyelinating oligodendrocytes in chronic lesions of multiple sclerosis. N. Engl. J. Med. 346, 165–173 (2002).

    Article  Google Scholar 

  32. Matsumoto, Y. et al. Differential proliferation rhythm of neural progenitor and oligodendrocyte precursor cells in the young adult hippocampus. PLoS One 6, e27628 (2011).

    Article  CAS  Google Scholar 

  33. Bellesi, M. Sleep and oligodendrocyte functions. Curr Sleep Med Rep. 1, 20–26 (2015).

    Article  Google Scholar 

  34. van Heyningen, P., Calver, A.R. & Richardson, W.D. Control of progenitor cell number by mitogen supply and demand. Curr. Biol. 11, 232–241 (2001).

    Article  CAS  Google Scholar 

  35. Draganski, B. et al. Neuroplasticity: changes in grey matter induced by training. Nature 427, 311–312 (2004).

    Article  CAS  Google Scholar 

  36. Scholz, J., Klein, M.C., Behrens, T.E. & Johansen-Berg, H. Training induces changes in white-matter architecture. Nat. Neurosci. 12, 1370–1371 (2009).

    Article  CAS  Google Scholar 

  37. Hu, Y. et al. Enhanced white matter tracts integrity in children with abacus training. Hum. Brain Mapp. 32, 10–21 (2011).

    Article  Google Scholar 

  38. Sampaio-Baptista, C. et al. Motor skill learning induces changes in white matter microstructure and myelination. J. Neurosci. 33, 19499–19503 (2013).

    Article  CAS  Google Scholar 

  39. Sagi, Y. et al. Learning in the fast lane: new insights into neuroplasticity. Neuron 73, 1195–1203 (2012).

    Article  CAS  Google Scholar 

  40. Ho, V.M., Lee, J.A. & Martin, K.C. The cell biology of synaptic plasticity. Science 334, 623–628 (2011).

    Article  CAS  Google Scholar 

  41. Kole, K. Experience-dependent plasticity of neurovascularization. J. Neurophysiol. 114, 2077–2079 (2015).

    Article  CAS  Google Scholar 

  42. Makinodan, M., Rosen, K.M., Ito, S. & Corfas, G. A critical period for experience-dependent oligodendrocyte maturation and myelination. Science 337, 1357–1360 (2012).

    Article  CAS  Google Scholar 

  43. Mangin, J.M., Li, P., Scafidi, J. & Gallo, V. Experience-dependent regulation of NG2 progenitors in the developing barrel cortex. Nat. Neurosci. 15, 1192–1194 (2012).

    Article  CAS  Google Scholar 

  44. Bergles, D.E. & Richardson, W.D. Oligodendrocyte development and plasticity. in Glia (eds. Barres, B.A., Freeman, M.R. & Stevens, B.) 139–165 (Cold Spring Harbor, 2015).

  45. Hines, J.H., Ravanelli, A.M., Schwindt, R., Scott, E.K. & Appel, B. Neuronal activity biases axon selection for myelination in vivo. Nat. Neurosci. 18, 683–689 (2015).

    Article  CAS  Google Scholar 

  46. Mensch, S. et al. Synaptic vesicle release regulates myelin sheath number of individual oligodendrocytes in vivo. Nat. Neurosci. 18, 628–630 (2015).

    Article  CAS  Google Scholar 

  47. Jolly, S., Fudge, A., Pringle, N., Richardson, W.D. & Li, H. Combining double fluorescence in situ hybridization with immunolabelling for detection of the expression of three genes in mouse brain sections. J. Vis. Exp. 109 10.3791/53976 (2016).

Download references


We thank our colleagues at University College London, especially S. Jolly, U. Grazini and L. Magno, for advice and reagents, and M. Grist and U. Dennehy for technical help. We thank M. Wegner (University of Erlangen, Germany) for the Sox10 antibody. This work was supported by the European Research Council (grant agreement 293544 to W.D.R.), the Wellcome Trust (100269/Z/12/Z to W.D.R.) and the Biotechnology and Biological Sciences Research Council (BB/L003236/1 to H.L.). L.X. was supported by the National Natural Science Foundation of China (grant 31471013). Pdgfra-CreERT2 mice can be obtained through with a material transfer agreement. Myrf loxP mice are available from Jackson Labs, strain 010607.

Author information

Authors and Affiliations



W.D.R. formed the hypotheses and obtained funding. I.A.M. adopted and developed the complex wheel test. B.E. provided MyrfloxP mice, advice and suggestions. W.D.R., I.A.M. and D.O. designed the experiments in Figure 1 and Supplementary Figure 1; D.O. and I.A.M. performed those experiments and DO analyzed the data. W.D.R., H.L. and L.X. designed all the other experiments and L.X. performed them, with assistance from A.S.-W., J.L.W. and A.D.F. H.L. identified Enpp6 and A.F. performed preliminary characterization. H.L. and W.D.R. supervised the work. W.D.R. wrote the paper with input from H.L., L.X. and B.E.

Corresponding author

Correspondence to William D Richardson.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Performance of P‑Myrf−/− versus P‑Myrf+/− mice on the complex wheel.

Three weeks after tamoxifen administration at P60-64 or P90-P94 (9), P‑Myrf−/− mice and their P‑Myrf+/− littermates [n=36 (20 males) and 32 (17 males) respectively] were housed singly in cages containing a complex wheel. Average wheel speeds were calculated for each 2-hour time window during the first seven nights (6pm-6am) and plotted as mean ± s.e.m (P‑Myrf−/−, red; P‑Myrf+/−, blue) were analyzed by two-way ANOVA with Bonferroni's post-hoc test. Each night was treated separately for multiple comparisons. *p < 0.05, **p < 0.01, ***p < 10−3, ****p < 10−4

[Night 1: 2 h, p=0.43, t=1.81; 4 h, p=0.029, t=2.83; 6 h, p=0.0089, t=3.20; 8 h, p=0.0044, t=3.40; 10 h, p=0.0017, t=3.66; 12 h, p=0.0036, t=3.46. Night 2: 2 h, p=0.0067, t=3.28; 4 h, p=0.0033, t=3.48; 6 h, p=0.063, t=2.57; 8 h, p=0.016, t=3.03; 10 h, p=0.11, t=2.37; 12 h, p=0.034, t=2.79. Night 3: 2 h, p=0.0019, t=3.63; 4 h, p=0.0041, t=3.43; 6 h, p=0.020, t=2.95; 8 h, p=0.072, t=2.53; 10 h, p>0.99, t=1.05; 12 h, p=0.24, t=2.06. Night 4: 2 h, p=0.020, t=2.96; 4 h, p=0.0010, t=3.80; 6 h, p=0.0007, t=3.90; 8 h, p=0.020, t=2.96; 10 h, p=0.033, t=2.79; 12 h, p=0.30, t=1.96. Night 5: 2 h, p=0.0006, t=3.94; 4 h, p=0.0052, t=3.36; 6 h, p=0.015, t=3.04; 8 h, p=0.13, t=2.32; 10 h, p=0.59, t=1.66; 12 h, p=0.17, t=2.42. Night 6: p<0.0001, t=4.42; 4 h, p<0.0001, t=4.42; 6 h, p<0.0001, t=4.46; 8 h, p=0.0046, t=3.39; 10 h, p=0.020, t=2.96; 12 h, p=0.095, t=2.42. Night 7: 2 h, p<0.0001, t=4.65; 4 h, p<0.0001, t=4.39; 6 h, p=0.0001, t=4.32; 8 h, p=0.0004, t=4.05; 10 h, p=0.20, t=2.13; 12 h, p=0.097, t=2.42. Degrees of freedom=396 throughout.]

Supplementary Figure 2 Two populations of high- and low-expressing Enpp6+ cells in vivo.

(a,b) Coronal sections of P60 mouse forebrains were incubated with an Enpp6 ISH probe and the signal developed using the NBT/BCIP method (Methods). (a) At a relatively long development time (150 minutes) two populations of Enpp6-positive cells are detected in the subcortical white matter – a few strongly-labeled large cell bodies (arrows) against a background of more numerous, small, weakly-labeled cells (arrowheads), consistent with the RNA-seq data27 (Fig. 3A) and the likelihood that the strongly-labeled cells are newly-differentiating oligodendrocytes and the weakly-labeled cells more mature, myelinating oligodendrocytes. (b) Reducing the development time to 60 minutes allows the strongly-labeled cells to be visualized preferentially (arrows). Images are representative of >3 similar experiments. Scale bar, 50 μm.

Supplementary Figure 3 Enpp6 expression is oligodendrocyte-specific.

Forebrain sections of P90 mice (caged without wheels) were analyzed by ISH for Enpp6 mRNA followed by immuno-fluorescence labeling for stage-specific markers of the oligodendrocyte lineage (Sox10, Olig2, CC1). Alternatively, sections were analyzed by double fluorescence ISH for Enpp6 and Pdgfra. (a,b) All Enpp6+ cells were also Olig2+ and Sox10+. (c) No Enpp6+ cells were Pdgfra+ OPs. (d) Most Enpp6+ cells were CC1+ mature or maturing oligodendrocytes (Motor cortex, 98.2 ± 1.8%; Subcortical white matter, 86.9% ± 3.8%) (Supplementary Table 1). Images are representative of >3 similar experiments. Scale bar, 25 μm.

Supplementary Figure 4 The role of novel motor activity in stimulating OP differentiation.

OPs are continuously cycling in the young adult CNS in response to mitogenic growth factors such Pdgf. After division one or both daughter OPs can rest in the G1 phase of the cell cycle, sometimes for days or weeks, before either differentiating or entering another division cycle3,4,7,9,31. Our data suggest that electrical activity in axon(s) cans stimulate OPs that are paused in G1 to differentiate - losing Pdgfra expression and expressing Enpp6, Mbp and other myelin gene products instead. The newly-differentiating oligodendrocytes have a distinctive spidery morphology in vivo; they remain like this for several days in vivo in rodents31 before down-regulating Enpp6 (faint pink line) and assuming the typical morphology of mature myelinating oligodendrocytes. The Enpp6high early-differentiating oligodendrocytes and Enpp6low myelinating oligodendrocytes probably contribute to improving circuit performance in the early and late stages of motor learning, respectively.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 and Supplementary Table 1 (PDF 1064 kb)

Supplementary Methods Checklist (PDF 696 kb)

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xiao, L., Ohayon, D., McKenzie, I. et al. Rapid production of new oligodendrocytes is required in the earliest stages of motor-skill learning. Nat Neurosci 19, 1210–1217 (2016).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing